Shuler M F, Tatti K M, Wade K H, Moran C P
Department of Microbiology and Immunology, Emory University, Atlanta, Georgia 30322, USA.
J Bacteriol. 1995 Jul;177(13):3687-94. doi: 10.1128/jb.177.13.3687-3694.1995.
We have examined the role of the most highly conserved region of bacterial RNA polymerase sigma factors by analyzing the effect of amino acid substitutions and small deletions in sigma E from Bacillus subtilis. sigma E is required for the production of endospores in B. subtilis but not for vegetative growth. Strains expressing each of several mutant forms of sigE were found to be deficient in their ability to form endospores. Single amino acid substitutions at positions 68 and 94 resulted in sigma factors that bind with less affinity to the core subunits of RNA polymerase. The substitution at position 68 did not affect the stability of the protein in B. subtilis; therefore, this substitution probably did not have large effects on the overall structure of the sigma factor. The substitution at position 68 probably defines a position in sigma E that closely contacts a subunit of RNA polymerase, while the substitution at position 94 may define a position that is important for protein stability or for binding to core RNA polymerase.
我们通过分析枯草芽孢杆菌中σE氨基酸替换和小片段缺失的影响,研究了细菌RNA聚合酶σ因子最高度保守区域的作用。σE是枯草芽孢杆菌产生芽孢所必需的,但对营养生长并非必需。表达几种sigE突变形式的菌株在形成芽孢的能力上存在缺陷。68位和94位的单个氨基酸替换导致σ因子与RNA聚合酶核心亚基的结合亲和力降低。68位的替换不影响该蛋白在枯草芽孢杆菌中的稳定性;因此,这种替换可能对σ因子的整体结构没有太大影响。68位的替换可能确定了σE中与RNA聚合酶一个亚基紧密接触的位置,而94位的替换可能确定了一个对蛋白质稳定性或与核心RNA聚合酶结合很重要的位置。